An asymmetric protoplast fusion and screening method for generating celeriac cybrids

Abstract Celeriac F1 hybrid seed production is currently complicated due to the instability of cytoplasmic male sterile lines. To develop alternative alloplasmic CMS lines, an asymmetric protoplast fusion and hybrid screening methodology was established. Celeriac suspension cells protoplasts were us...

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Autores principales:	Silvia Bruznican, Tom Eeckhaut, Johan Van Huylenbroeck, Ellen De Keyser, Hervé De Clercq, Danny Geelen
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/79be3ae4b55547fa8632286d773a1e53
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spelling	oai:doaj.org-article:79be3ae4b55547fa8632286d773a1e532021-12-02T13:33:44ZAn asymmetric protoplast fusion and screening method for generating celeriac cybrids10.1038/s41598-021-83970-y2045-2322https://doaj.org/article/79be3ae4b55547fa8632286d773a1e532021-02-01T00:00:00Zhttps://doi.org/10.1038/s41598-021-83970-yhttps://doaj.org/toc/2045-2322Abstract Celeriac F1 hybrid seed production is currently complicated due to the instability of cytoplasmic male sterile lines. To develop alternative alloplasmic CMS lines, an asymmetric protoplast fusion and hybrid screening methodology was established. Celeriac suspension cells protoplasts were used as the acceptor and carrot, coriander and white celery mesophyll protoplasts as the donor for protoplast fusion experiments. Acceptor cytoplasmic inheritance was inhibited by iodoacetamide treatment and donor nuclear genome inheritance was prevented by UV exposure. Protoplasts were selectively stained and fused using electroporation and polyethylene glycol, and candidate hybrid shoots were obtained. One chloroplast and three mitochondrial markers that could distinguish acceptor and donors organelles were used to characterize over 600 plants obtained after fusion events, without identifying any cybrid. In order to increase the testing efficiency a high number of micro plantlets were pooled and hence the presence of the carrot specific Atp1 marker in one of the pooled samples was detected. We demonstrated that fusion took place between celeriac and a carrot indicating that the creation of viable hybrids is possible although at a very low frequency. These findings open the path for new cytoplasmic hybridization and the isolation of novel CMS lines of celeriac.Silvia BruznicanTom EeckhautJohan Van HuylenbroeckEllen De KeyserHervé De ClercqDanny GeelenNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 11, Iss 1, Pp 1-12 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Silvia Bruznican Tom Eeckhaut Johan Van Huylenbroeck Ellen De Keyser Hervé De Clercq Danny Geelen An asymmetric protoplast fusion and screening method for generating celeriac cybrids
description	Abstract Celeriac F1 hybrid seed production is currently complicated due to the instability of cytoplasmic male sterile lines. To develop alternative alloplasmic CMS lines, an asymmetric protoplast fusion and hybrid screening methodology was established. Celeriac suspension cells protoplasts were used as the acceptor and carrot, coriander and white celery mesophyll protoplasts as the donor for protoplast fusion experiments. Acceptor cytoplasmic inheritance was inhibited by iodoacetamide treatment and donor nuclear genome inheritance was prevented by UV exposure. Protoplasts were selectively stained and fused using electroporation and polyethylene glycol, and candidate hybrid shoots were obtained. One chloroplast and three mitochondrial markers that could distinguish acceptor and donors organelles were used to characterize over 600 plants obtained after fusion events, without identifying any cybrid. In order to increase the testing efficiency a high number of micro plantlets were pooled and hence the presence of the carrot specific Atp1 marker in one of the pooled samples was detected. We demonstrated that fusion took place between celeriac and a carrot indicating that the creation of viable hybrids is possible although at a very low frequency. These findings open the path for new cytoplasmic hybridization and the isolation of novel CMS lines of celeriac.
format	article
author	Silvia Bruznican Tom Eeckhaut Johan Van Huylenbroeck Ellen De Keyser Hervé De Clercq Danny Geelen
author_facet	Silvia Bruznican Tom Eeckhaut Johan Van Huylenbroeck Ellen De Keyser Hervé De Clercq Danny Geelen
author_sort	Silvia Bruznican
title	An asymmetric protoplast fusion and screening method for generating celeriac cybrids
title_short	An asymmetric protoplast fusion and screening method for generating celeriac cybrids
title_full	An asymmetric protoplast fusion and screening method for generating celeriac cybrids
title_fullStr	An asymmetric protoplast fusion and screening method for generating celeriac cybrids
title_full_unstemmed	An asymmetric protoplast fusion and screening method for generating celeriac cybrids
title_sort	asymmetric protoplast fusion and screening method for generating celeriac cybrids
publisher	Nature Portfolio
publishDate	2021
url	https://doaj.org/article/79be3ae4b55547fa8632286d773a1e53
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An asymmetric protoplast fusion and screening method for generating celeriac cybrids

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